Patent Application
20210196970
None.
The present invention relates to systems and methods for treating cancer with cold atmospheric plasma.
Cholangiocarcinoma (CCA) is a rare and aggressive malignancy arising in the intrahepatic or extrahepatic biliary tract. It is often discovered in advanced late stages, and the prognosis is poor with a five-year survival rate under 20%. DeOlivcira, M. L., S. C. Cunningham., J. L. Cameron, et al., Cholangiocarcinoma: Thirty-one-year experience with 564 patients at a single institution. Ann Surg, 2007. 245 (5): p.755-'762; Horgan, A. M., E. Amir, T. Walter, et al., Adjuvant therapy in the treatment of biliary tract cancer: A systematic review and meta-analysis. J Clin Oncol, 2012. 30 (16): p. 1934- 40. CCAs are classified by location into intrahepatic cholangiocarcinoma (ICCA), perihilar cholangiocarcinoma (PHC), or distlil cholangiocarcinoma (DCCA) subtypes. Further ICCA has up to a 70% recurrence rate after surgical resection. Mazzaferro, V., A. Gorgen, S. Roayaie, et al., Liver resection and transplantation for intrahepatic cholangiocarcinoma. J Hepatol, 2020. 72 (2): p. 364-37. Surgical resection or liver transplantation at an early stage are the best options for curative treatment of CCA. Shen, W. F., W. Zhong, P. Xu, et al., Clinicopathological and prognostic analysis of 429 patients with intrahepatic cholangiocarcinoma. World J Gastroenterol, 2009. 15 (47): p. 5976-82. Chemoresistance presents a challenge in administering adjuvant chemotherapy in all classification types and as a result, CCA is known for poor clinical outcomes. Marin, J. J. G., E. Lozano, E. Herraez, et al., Chemoresistance and chemosensitization in cholangiocarcinoma. Biochim Biophys Acta Mol Basis Dis, 2018. 1864 (4 Pt B): p. 1444- 1453; Kirstein, M. M. and A. Vogel, Epidemiology and risk factors of cholangiocarcinoma. Vise Med, 2016. 32 (6): p. 395-400.
For patients with recurrent CCA, gemcitabine and fluorouracil (5-FU) have been standard options as individual treatments or drug combination therapy for years. Cisplatin plus gemcitabine versus gemcitabine for biliary tract cancer. New England Journal of Medicine, 2010; Hezel, A. F. and A. X. Zhu, Systemic therapy for biliary tract cancers. Oncologist, 2008. 13 (4): p. 415-23; Penz, M., G. V. Kornek, M. Raderer, et al., Phase ii trial of two-weekly gemcitabine in patients with advanced biliary tract cancer. Ann Oncol, 2001, 12 (2): p183-6. The FOLFIRINOX protocol is a drug regimen consisting of fluorouracil (5-FU), leucovorin, irinotecan, and oxaliplatin. This novel regimen emerged as a first line therapy in pancreatic cancers, but it is not yet standard clinical practice for CCA. Lambert, A., C. Gavoille, and T. Conroy, Current status on the place of folfirinox in metastatic pancreatic cancer and future directions. Therap Adv Gastroenterol, 2017, 10 (8): p. 631-645. A small retrospective study, including 32 patients with CCA, had a patient tailored approach to FOLFIRINOX regimen and a disease control rate of 76%. Ulusakarya, A., A. Karaboue, 0. Ciacio, et al., A retrospective study of patient-tailored folfirinox as a first-line chemotherapy for patients with advanced biliary tract cancer. BMC Cancer, 2020. 20 (1): p. 515. In a phase two-three clinical trial, FOLFIRINOX increased overall survival over gemcitabine treatment from 6.8 to 11.1 months in patients with metastatic pancreatic cancer. Conroy, T., F. Desseigne, M. Ychou, et al., Folfirinox versus gemcitabine for metastatic pancreatic cancer. N Engl. I Med, 2011. 364 (19): p. 1817-1825. However the regimen was not well tolerated; the incidence of thrombocytopenia, neutropenia, and febrile neutropenia were significantly higher in FOLFIRINOX treatment patients. To address this limitation, studies have focused on reducing dose or modifying the four components. Dosage iterations of modified FOLFOX-4, FOLFOX-5, and FOLFOX-7 have been used to treat pancreatic, colorectal, and bladder cancers. Dodagoudar, C., D. C. Doval, A. Mahanta, et al., Folfox-4 as second-line therapy after failure of gemcitabine and platinum combination in advanced gall bladder cancer patients. Jpn J Clin Oncol, 2016. 46 (1): p. 57-62; Schinzari, G. E. Rossi, G. Mambella, et al., First-line treatment of advanced biliary ducts carcinoma: A randomized phase ii study evaluating 5-fu/lv plus oxaliplatin (folfox 4) versus 5-fu/lv (de gramont regimen). Anticancer Res, 2017. 37 (9): p. 5193-5197; Conroy, T., P. Hammel, M. I-lebbar, et al., Folfirinox or gemcitabine as adjuvant therapy for pancreatic cancer. N Engl. I Med, 2018. 379 (25): p. 2395-2406.
Cold atmospheric plasma (CAP) has been extensively studied in various biomedical fields. It is a novel approach to targeted cancer treatment and has demonstrated its anti-cancer effects in vitro. See, Rowe, W., X. Cheng, L. Ly, et al., The Canady Helios cold plasma scalpel significantly decreases viability in malignant solid tumor cells in a dose dependent manner. Plasma, 2018. 1 (1): p. 177-188; Barekzi, N. and M. Laroussi, Effects of low temperature plasmas on cancer cells. Plasma Processes and Polymers, 2013. 10 (12): p. 1039-1050; Barekzi, N. and M. Laroussi, Dose-dependent killing of leukemia cells by low to temperature plasma. Journal of Physics D: Applied Physics, 2012. 45 (12); and Keidar, M., R. Walk, A. Shashurin; et al., Cold plasma selectivity and the possibility of a paradigm shift in cancer therapy. Br J Cancer, 2011. 105 (9): p. 1295-301. The detailed mechanism has not been fully elucidated; however, studies have established that CAP selectively induces apoptosis and DNA damage in tumor cells. Arndt, S., M. Landthaler, J. L. Zimmermann, et al., Effects of cold atmospheric plasma (cap) on ss-defensins, inflammatory cytokines, and apoptosis-related molecules in keratinocytes in vitro and in vivo. PLoS One, 2015. 10 (3): p. e0120041; Bauer, G., D. Sersenova, D. B. Graves, et al., Cold atmospheric plasma and plasma activated medium trigger rans-based tumor cell apoptosis. Sci Rep, 2019. 9 (1): p. 14210; Cheng, X., W. Rowe, L. Ly, et al., Treatment of triple-negative breast cancer cells with the Canady cold plasma conversion system: Preliminary results. Plasma, 2018. 1 (1): p. 218-228. Further research indicates low doses of CAP does not damage normal tissue. See, e.g., Lee, J. II., J. Y. Om, Y. H. Kim, et al., Selective killing effects of cold atmospheric pressure plasma with no induced dysfunction of epidermal growth factor receptor in oral squamous cell carcinoma. PLoS One, 2016. 11 (2): p. e0150279. Recently, indirect CAP treatment was effective for the treatment of CCA in vitro, selectively killing CCA cells over normal hepatocytes. Vaquero, J., F. Judee, M. Vallette, et al., Cold-atmospheric plasma induces tumor cell death in preclinical in vivo and in vitro models of human cholangiocarcinoma. Cancers, 2020. 12 (5). Research on CAP in combination with other therapies has shown some potential synergism with anti-neoplastic agents in melanoma cells (Sagwal, S. K., G. Pasqual-Melo, Y. Bodnar, et al., Combination of chemotherapy and physical plasma elicits melanoma cell death via upregulation of s1c22a1 6. Cell Death Dis, 2018. 9 (12): p. 1179), drug loaded nanoparticles in breast cancer cells (Zhu, W., S. J. Lee, N. J. Castro, et al., Synergistic effect of cold atmospheric plasma and drug loaded core-shell nanoparticles on inhibiting breast cancer cell growth. Sci Rep, 2016. p. 21974), and gemcitabine in murine pancreatic cancer cells (Masur, K., M. van Behr, S. Bekeschus, et al., Synergistic inhibition of tumor cell proliferation by cold plasma and gemcitabine. Plasma Processes and Polymers, 2015.12 (12): p. 1377-1382).
Delivery of cold atmospheric plasma at the surgical margins immediately after tumor resection has shown potential as an anti-cancer therapy. A Canady Cold Plasma Conversion System is an electrosurgical system that produces CAP for the treatment of surgical margins upon tumor resection (U.S. Patent No. 9,999,462). One of the advantages of cold atmospheric plasma systems is that the CAP temperature remains between 26-30 ° C. during the duration of the treatment (Cheng, X., et al., Treatment of Triple-Negative Breast Cancer Cells with the Canady Cold Plasma Conversion System: Preliminary Results. Plasma, 2018. 1 (1): p. 218-228) and does not cause any thermal or physical damage to normal tissue (Ly, L., et al., A New Cold Plasma Jet: Performance Evaluation of Cold Plasma, Hybrid Plasma and Argon Plasma Coagulation. Plasma, 2018. 1 (1): p. 189-200).
Cholangiocarcinoma (CCA) is a rare biliary tract cancer with a low five-year survival rate and high recurrence rate after surgical resection. Currently treatment approaches include systemic chemotherapeutics such as FOLFIRINOX, a chemotherapy regimen is a possible treatment for severe CCA cases. A limitation of this chemotherapy regimen is its toxicity to patients and adverse events. There exists a need for therapies to alleviate the toxicity of a FOLFIRINOX regimen while enhancing or not altering its anticancer properties. Cold Atmospheric Plasma (CAP) is a technology with a promising future as a selective cancer treatment. In this study, FOLFIRINOX treatment alone at the highest dose tested (53.8 nM fluorouracil, 13.1 nM Leucovorin, 5.1 nM irinotecan, and 3.7 nM Oxaliplatin) reduced CCA cell viability to below 20% while CAP treatment alone for 7 min reduced viability to 3% (p<0.05). An analysis of cell viability, proliferation, and cell cycle demonstrated that CAP in combination with FOLFIRINOX is more effective than either treatment alone at a lower FOLFIRINOX dose of 6.73 nM fluorouracil, 1.71 nM leucovorin, 0.63 nM irinotecan, and 0.47 nM oxaliplatin and a shorter CAP treatment of 1, 3, or 5 minutes. CAP reduces the toxicity burden of FOLFIRINOX. FOLFIRINOX and CAP at various dose levels to quantify changes in cell viability and cell cycle progression. FOLFIRINOX administered as a first line therapy followed by CAP treatment produces an in vitro synergistic effect.
In a preferred embodiment, the present invention is a method for treatment of cholangiocarcinoma with cold atmospheric plasma and Folfirinox. The method comprises pre-operatively treating a patient having a cholangiocarcinoma with low-dosage FOLFIRINOX not exceeding 26.91 nM fluorouracil, 6.84 nM leucovorin, 2.53 nM irinotecan, and 1.87 nM oxaliplatin, surgically removing the cholangiocarcinoma, applying cold atmospheric plasma to the surgical margins surrounding the area in the patient from which the tumor was removed, and treating the patient with low dosage FOLFIRINOX post-operatively. The method further may include treating the patient with FOLFIRINOX intra-operatively. The low-dosage FOLFIRINOX has at least 6.73 nM fluorouracil, 1.71 nM leucovorin, 0.63 nM irinotecan, and 0.47 nM oxaliplatin. In another embodiment the low-dosage FOLFIRINOX has no more than 13.45 nM fluorouracil, 3.42 nM leucovorin, 1.26 nM irinotecan, and 0.94 nM oxaliplatin.
Still other aspects, features, and advantages of the present invention are readily apparent from the following detailed description, simply by illustrating a preferable embodiments and implementations. The present invention is also capable of other and different embodiments and its several details can be modified in various obvious respects, all without departing from the spirit and scope of the present invention. Accordingly, the drawings and descriptions are to be regarded as illustrative in nature, and not as restrictive. Additional objects and advantages of the invention will be set forth in part in the description which follows and in part will be obvious from the description or may be learned by practice of the invention.
For a more complete understanding of the present invention and the advantages thereof, reference is now made to the following description and the accompanying drawings, in which:
Cholangiocarcinomas are rare with a low five-year survival rate. Cold atmospheric plasma (CAP) is a promising technology as a selective cancer treatment due to its anticancer properties. In pancreatic and liver cancers, FOLFIRINOX has emerged as an effective combination cancer drug treatment. It has been reported that FOLFIRINOX increased overall patient survival over gemcitabine treatment in patients with metastatic pancreatic cancer but is limited due to toxicity. See, T. Conroy et al., N. Engl. J. Med., 364, pp. 1817-1825 (2011) and T. Conroy et al., N. Engl. J. Med., 379, pp. 2395-2406 (2018).
A method for treating cancer with a combination of FOLFIRINOX and CAP in accordance with a preferred embodiment of the present invention is shown in
A preferred embodiment of a CAP enabled generator is described with reference to the drawings. A gas-enhanced electrosurgical generator 200 in accordance with a preferred embodiment of the present invention is shown in
A generator housing front panel 210 is connected to the housing 202. On the face front panel 210 there is a touchscreen display 212 and there may be one or a plurality of connectors 214 for connecting various accessories to the generator 200. For a cold atmospheric plasma generator such as is shown in
As shown in
As shown in
Another embodiment, shown in
In the above-disclosed embodiment, a cold atmospheric plasma below 35° C. is produced. When applied to the tissue surrounding the surgical area, the cold atmospheric plasma induces metabolic suppression in only the tumor cells and enhances the response to the drugs that are injected into the patient.
The cold plasma applicator 500 may be in a form such as is disclosed in U.S. Pat. No. 10,405,913 and shown in
The intrahepatic poorly differentiated cholangiocarcinoma cell line, KKU-055, was purchased from Sekisui XenoTech, LLC (Kansas City, Kans.). Cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum and 1% Pen Strep (Thermo Fisher Scientific, Waltham, Mass., USA). Cells were lifted with Trypsin-EDT A and seeded in 12-well plates at 100,000 cells/well or 50,000 cells/well in 1 mL complete media. Cells were then incubated 24 hours at 37° C. and 5% CO2 prior to drug or CAP treatment. All experiments were performed at the Jerome Canady Research Institution for Advanced Biological and Technological Sciences (JCRI-ABTS) in Takoma Park, Md., USA.
All CAP treatments were generated with a US Medical Innovations LLC 22-601 MCa high frequency electrosurgical generator, a Canady Helios™ Cold Plasma System, paired with a Canady Helios Cold Plasma™ Scalpel. All CAP tests were conducted with a constant helium flow rate of 3 L/min, at a power setting of 120 p, which corresponds to 28.7 W. Treatment durations were up to 7 minutes. The distance between the tip of the plasma scalpel and media surface was constant at 1.5 cm. Immediately after CAP treatment, cells were transferred to a 37° C. and 5% CO2 humidified incubator and cultured up to 72 hours.
The four FOLFIRINOX drugs were individually diluted in DMSO then combined in a stock solution at the clinical dose ratio of oxaliplatin (Sigma Aldrich #PHR 1528) 85 mg/m2, leucovorin (Sigma Aldrich #PI JR 1541) 400 mg/1112, irinotecan (Sigma Aldrich #11406) 180 mg/m2, and 5-fluorouracil (Sigma Aldrich #PHR 1227) 400 mg/1112. FOLFIRINOX doses will be referred to by their corresponding concentration of 5-flourouracil [5-FU]. Further dilutions of the four drugs into the FOLFIRINOX mix were made with complete cell culture media. Cells were incubated with a single dose of drug treatment for 24 hours prior to CAP treatment.
Cellular viability and proliferation were assessed through a Thiazolyl Blue Tetrazolium Bromide (MTT, Abcam ab146345) assay performed 48 hours after CAP treatment. Cells were incubated with MTI solution for 3 hours at 37 and 5% CO2 humidified incubator. The absorbance of the dissolved compound was measured by BioTek Synergy I ITX (Winooski, Vt., USA) microplate reader at 570 nm. Viability assays were repeated at least 3 times with a minimum of 2 intra experimental replicates. For each assay cell viability was calculated by normalizing non-treated cells.
Confocal microscopy analysis was prepared in the following manner. One round platinum lined cover glass 12 mm in diameter was placed in each well of a 12-well plate then coated with fibronectin and collagen II for at least 12 hours. Cells were then seeded on cover glass inside of wells to normalize treatment to MTT assays and IncuCyte analysis. After selected drug treatment, CAP treatment, or combination treatment cultures were fixed with ice cold (−20° C.) methanol for 10 minutes. Then cells were stained with Alexa Fluor 488-conjugated Ki-67 Rabbit mAh (Cell Signaling Technology, #11882) or isotype control (Cell Signaling Technology, #4340) antibodies according to Immunofluorescence General Protocol by Cell Signaling Technology (Danvers, Mass., USA). Cells were incubated overnight at 4° C. protected from light. The cover slides were then carefully moved onto glass slides and covered with Anti fade Mounting Reagent with DAPI (Vector Laboratories, H-1500) drops and then a 1 mm cover slide. The slides were allowed to cure for up to 2 nights in a 4° C. refrigerator then sealed with clear nail polish.
Images were taken with Zeiss Confocal 510 LSM (Oberkochen, Germany), analyzed with Zeiss ZenLite (2012) software, and Ki-67 positivity was calculated in Microsoft Excel 2019 (Redmond, Wash., USA).
Cell cycle phase contrast images were collected on the IncuCyte® Live-Cell Analysis System (Essen Bioscience, Ann Arbor, Mich.). A stable KKU-055 cell line was established through 5 μg/mL puromycin (Sigma Aldrich P8833) selection after transfection with the IncuCyte® Red/Green Lentivirus Reagent (IncuCyte #4779) for labelling and indication of in vitro cell cycle. Red indicated G1 phase and Green indicated S/G2/M phases while unlabeled cells indicated M-G1 transition phase or dead cells. In-vitro cell growth images were collected at 1-hour intervals up to 72 hours after each treatment condition. The percent of cell confluence and detailed cell counts per well were quantified by the IncuCyte® Cell by Cell Analysis then plotted in Microsoft Excel 2019.
Data was plotted by Microsoft Excel 2019 as mean±standard error of the mean. Student unpaired t-tests and two-way analysis of variance (ANOVA) were used to determine significant differences between the groups. Significant CAP-drug combination effects were followed by post hoc tests with Bonferroni correction. To determine significance of independent and combined treatment groups with p-value<0.05 considered statistically significant.
To determine the possible synergistic effects of FOLFIRINOX on KKU-055 cells, an optimal dosage of the four drugs in combination must be able to reduce cell viability significantly. A serial dilution of 6 doses of FOLFIRINOX was done to establish a baseline toxicity measurement for each dose (Table 1 and
Table 1 shows drug concentrations of the four FOLFIRINOX components for each dose level is the serial dilution. This corresponds to the [5-FU] notation in
A dose dependence experiment was performed on KKU-055 cells to establish CAP efficacy. MTT assays were conducted 48 hours post CAP treatment. Cell viability was significantly reduced by CAP for all durations, and the highest treatment of 7 minutes reduced viability to 3% (p<0.005). FIG. 8 is a bar graph illustrating a reduction of KKU-055 cell viability 48 hours after CAP treatment for 1-7 minutes at 120 p which corresponds to 28.7 W compared to untreated controls (cohort =4, 2/cohort, n=8/test), *P<0.05.
KKU-055 cells were exposed to 24 hours of FOLFIRINOX pretreatment at 6.7-53.8 nM [5-FU] (Table I) and CAP at 120 p for 1, 3, or 5 minutes. Viability reduction was measured 48 hours after treatment (
A two-way ANOVA test followed by post hoc Fisher exact tests (with Bonferroni correction) was conducted on this combination treatment experiment. Sources of variation were a change in either CAP dose or FOLFIRINOX dose. Then the variance between the two was tested to determine if one treatment had an effect of the other. There were three hypotheses for this test; H1: The observed viability between drug dosage groups is equal; H2: the observed viability between CAP dosage groups is equal; and H3: there is no interaction between the two treatments. For all three hypotheses p<0.05, so we can reject each one. Student paired t-tests and two-way ANOVA test followed by post hoc Fisher exact tests (with Bonferroni correction) were then conducted to compare each combination treatment with every other experiment group (Table 2).
Dosage combinations were considered synergetic when combination treatment reduced viability significantly more than the corresponding CAP or FOLFIRINOX dosage alone. In cases when the FOLFIRINOX dose was 13.5 nM [5-FU] or higher the drug alone was strong enough to reduce KKU-055 viability to below 30%, and this made drug treatment significantly more effective than 1 or 3 minutes of CAP (cohort=4, 2/cohort, n=8 t test p<0.05,
Table 2 is a chart showing the comparison of the reduction of viability between treatment groups. Whether there is statistical difference p<0.005 and if that difference is extremely significant p<1×10−5(Student's t test with Bonferroni's correction).
Cell proliferation was examined by Ki-67/DAPI co-staining at 6, 24, or 48 hours post CAP, FOLFIRINOX, or combination treatment. The 6.7 nM 5-FU dose of drug (Table 1) was combined with 1, 3, and 5 minutes of CAP. In five images, nuclei that were in focus were outlined and each mean fluorescence intensity (MFI) of Ki-67 channel was recorded. The mean of Ki-67 MFI was calculated for each treatment group including for No Treatment and Isotype control. A Ki-67+cell threshold was determined as a cell with an MFI greater than the lowest mean of MFI of all groups other than Isotype control. There was a significant (cohort=3, 2/cohort, n=6, t test p<0.05) decrease in cell count with FOLFIRINOX and 3 minutes of CAP treatment combined at 6 hours compared to no treatment controls (
KKU-055 cells were imaged 6, 24, and 48 hours after CAP or CAP and FOLFIRINOX treatments with an untreated negative control.
Experiments were designed to measure cell confluence and cell cycle distribution after combining the 6.7 nM 5-FU dose of FOLFIRINOX (Table 1) and CAP at 3 and 5 minutes. Cells were placed in the IncuCyte Live Cell imaging system immediately after CAP where confluence was monitored.
Images of 0 hours, 24 hours, and 48 hours timepoints demonstrates cell confluence within treatment wells. In the images, morphological differences were seen between experiment conditions. No treatment and drug only treated cells were confluent at 48 hours with most cells visibly fluorescent. In combination treatment wells, cells were not confluent and large clusters of cellular debris was visible after 48 hours of treatment.
The number of cells in different phases of the cell cycle was quantified through fluorescence measurements. The quantifications during the first 48 hours after treatment are shown at the CAP 1, 3, and 5-minute doses (
Cholangiocarcinoma treatment research aims to improve available chemotherapeutic options and FOLFIRINOX is promising as a novel, effective, yet toxic treatment. A clinical goal now is to establish a standard FOLFIRINOX dosage based on clinical trials. Multiple phase 1 and 2 studies are underway with encouraging results for FOLFIRINOX treatment in different doses over gemcitabine plus cisplatin, however there is no standard. The early issues in these studies are toxicity of FOLFIRINOX and early triumphs show that the regimen can be safe in patients able to tolerate it. These trials attempt to minimize toxicities by reducing or modifying drug doses because patients are excluded due to low performance status. See, Dodagoudar, C., D. C. Doval, A. Mahanta, et al., Folfox-4 as second-line therapy after failure of gemcitabine and platinum combination in advanced gall bladder cancer patients. Jpn J Clin Oncol., 2016. 46 (1): p. 57-62; and Funasaka, C., Y. Kanemasa T. Shimoyama, et al., Modified folfox-6 plus bevacizumab chemotherapy for metastatic colorectal cancer in patients receiving hemodialysis: A report of three cases and review of the literature. Case Rep Oncol, 2019. 12 (2): p, 657-665.
CAP is a promising therapy for CCA because of its selectivity of cancer cells in bile duct, liver, and pancreatic cases in vitro. However systemic risks have not been extensively studied in clinical cases due to limited CAP use on patients. The lack of severe side effects in one cohort of 20 patients with oral cancer is encouraging. CAP has already been studied in vitro and in vivo with gemcitabine treatment, a standard option in CCA and pancreatic cancer regimens. Masur, K., M. van Behr, S. Bekeschus, et al., Synergistic inhibition of tumor cell proliferation by cold plasma and gemcitabine. Plasma Processes and Polymers, 2015, 12 (12): p. 1377-1382. Liedtke, K. R., E. Freund, M. Hermes, et al., Gas plasma-conditioned ringer's lactate enhances the cytotoxic activity or cisplatin and gemcitabine in pancreatic cancer in vitro and in vivo. Cancers (Basel), 2020. 12 (1); Brulle, L., M. Vandamme, D. Ries, et al., Effects of a non-thermal plasma treatment alone or in combination with gemcitabine in a mia paca2-luc orthotopic pancreatic carcinoma model, PLoS One, 2012. 7 (12): p. e52653.
These reports support a combined anti-tumor effect, demonstrating that CAP has potential to increase anti-tumor effectiveness of current medicines. Recently, cold atmospheric plasma has also been studied in combination with other treatments to establish potential synergetic therapy. In this study, CAP was combined with a FOLFIRINOX regimen to treat cholangiocarcinoma cells as there exists a need to examine interactions between CAP and novel chemotherapeutics.
This study demonstrates that both CAP and FOLFIRINOX individually and in combination effectively reduce cell viability suggesting that FOLFIRINOX dosage can be reduced if paired with CAP for the treatment or CCA. Synergy was seen through MTT assays at various doses of FOLFIRINOX and CAP (Table 2). Confocal microscopy and IncuCyte imaging demonstrated a decrease in cell counts and changes in cell morphology after treatment which was consistent with the reduction in viability shown in
This is the first study to investigate the synergistic interaction between CAP and FOLFIRINOX for the treatment of cholangiocarcinoma. Our finding of synergism between CAP and chemotherapeutics has great potential. CAP and FOLFIRINOX can be combined to reduce cholangiocarcinoma tumor cell viability and proliferation. We determined the dosage combinations in which viability reduction could be enhanced by adding 1-5 minutes of low temperature plasma to a very low dose of FOLFIRINOX (6.73 nM fluorouracil, 1.71 nM leucovorin, 0.63 nM irinotecan, and 0.47 nM oxaliplatin). A combination therapy would be advantageous for patients where an intense FOLFIRINOX regimen is too aggressive, and this warrants further clinical research. We focused on the low doses of FOLFIRINOX to reduce overall chemotherapeutic burden in vitro as a model of lower toxicity in vivo. If a lower dose of FOLFIRINOX is administered, patients with low performance status can have more treatment options. Knowledge or the interactions between CAP and chemotherapeutics is of clinical value and can lead to more personalized medicine and a lower chemotherapy burden on patients in the future.
The effectiveness of Canady Helios™ Cold Atmospheric Plasma in combination with a FOLFIRINOX regimen was explored. We found that a combination treatment can be significantly more effective than either CAP or FOLFIRINOX alone in reducing cholangiocarcinoma cell viability. We are the first to demonstrate the in vitro synergistic effect of a FOLFIRINOX treatment and CAP, and our data suggests CAP could be a possible adjuvant therapy for cholangiocarcinoma. It is important that CAP alone can selectively induce tumor cell death, however our results demonstrate that CAP can potentially reduce the dose of chemotherapeutic drugs needed for cancer patients. Future studies may examine the cellular pathways involved in these synergistic characteristics and identify the ideal dose of treatment that has the lowest feasible toxicity with the most productive outcome. This study provides insights for the clinical application of CAP for cholangiocarcinoma cancer treatment.
The foregoing description of the preferred embodiment of the invention has been presented for purposes of illustration and description. It is not intended to be exhaustive or to limit the invention to the precise form disclosed, and modifications and variations are possible in light of the above teachings or may be acquired from practice of the invention. The embodiment was chosen and described in order to explain the principles of the invention and its practical application to enable one skilled in the art to utilize the invention in various embodiments as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the claims appended hereto, and their equivalents. The entirety of each of the aforementioned documents is incorporated by reference herein.
The present application claims the benefit of the filing date of U.S. Provisional Patent Application Ser. No. 62/953,783 filed by the present inventors on Dec. 26, 2019. The aforementioned provisional patent application is hereby incorporated by reference in its entirety.
| Number | Date | Country | |
|---|---|---|---|
| 62953783 | Dec 2019 | US |
